Abstract
BACKGROUND: We have developed a semi-automated fluorometric method that utilizes ethidium bromide (EB), a common substrate of bacterial efflux pumps. The method is sufficiently sensitive to characterize the efflux pump systems of bacteria. Because EB is also recognized and extruded by ATP-binding cassette (ABC) transporters and these have similarity to P-glycoprotein (P-gp), the method has been extended for the evaluation of agents that can inhibit the extrusion of EB on a real-time basis by mouse lymphoma cells containing the human ABCB1 (mdr1) gene.
MATERIALS AND METHODS: Monitoring of uptake and extrusion of EB was assessed using the Rotor-Gene 3000 (Corbett Research) under different conditions.
RESULTS: Whereas extrusion of EB took place readily, the addition of known inhibitors of efflux pumps (verapamil, reserpine) caused retention of EB.
CONCLUSION: This method is inexpensive and allows the detection of neoplastic cells with increased efflux activity as well as the screening of large numbers of compounds for inhibition of the P-gp.
| Original language | English |
|---|---|
| Pages (from-to) | 2173-2177 |
| Number of pages | 5 |
| Journal | Anticancer Research |
| Volume | 29 |
| Issue number | 6 |
| Publication status | Published - Jun 2009 |
Keywords
- Adrenergic Uptake Inhibitors
- Animals
- Automation
- Biological Transport
- Calcium Channel Blockers
- Cell Survival
- Drug Resistance, Multiple
- Drug Resistance, Neoplasm
- Ethidium
- Fluorometry
- Humans
- Lymphoma, T-Cell
- Mice
- P-Glycoprotein
- P-Glycoproteins
- Reserpine
- Verapamil
- Journal Article
- Research Support, Non-U.S. Gov't
